维生素D对高糖诱导足细胞胰岛素抵抗的保护作用

临床儿科杂志 ›› 2014, Vol. 32 ›› Issue (5): 471-475.

维生素D对高糖诱导足细胞胰岛素抵抗的保护作用

邹敏书¹, 余健¹, 聂国明¹, 罗莉漫¹, 徐洪涛¹, 尹晓玲², 周建华²

1. 广州军区武汉总医院儿科（湖北武汉　430070）；2. 华中科技大学同济医学院同济医院儿科（湖北武汉　430030）

收稿日期:2013-12-07 出版日期:2014-05-15 发布日期:2014-05-15

Protective effect of vitamin D on high glucose-induced podocyte insulin resistance

ZOU Minshu¹, YU Jian¹, NIE Guoming¹, LUO Liman¹, XU Hongtao¹, YIN Xiaoling², ZHOU Jianhua²

(1. Department of pediatrics, Wuhan General Hospital of Guangzhou Command of PLA, Wuhan 430070，China; 2. Department of Pediatrics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China)

Received:2013-12-07 Online:2014-05-15 Published:2014-05-15

摘要/Abstract

摘要：

足细胞+5mmol/L葡萄糖（A组），足细胞+5mmol/L葡萄糖+1nmol/L丙二醇（B组），足细胞+30mmol/L葡萄糖+1nmol/L丙二醇（C组），足细胞+30mmol/L葡萄糖+1nmol/L丙二醇+1nmol/L VitD（D组）。培养48 h测定足细胞凋亡百分率，MTT法测定足细胞活性，RT-PCR测定足细胞维生素D受体（VDR）及胰岛素受体底物蛋白1（IRS1）的表达，Western blotting检测IRS1、肌醇依赖蛋白激酶（Akt）、细胞外调节蛋白激酶（ERK1/2）的表达及其磷酸化水平。结果　四组间足细胞凋亡及活力，VDR和IRS1基因表达水平，信号蛋白IRS1、ERK_1/2磷酸化水平的差异均有统计学意义（P均<0.05），Akt磷酸化水平的组间差异无统计学意义（P>0.05）。与A、B和D组相比，C组足细胞凋亡百分率升高，足细胞活力降低，VDR和IRS1 mRNA表达下调，IRS1磷酸化水平下降，ERK_1/2磷酸化水平升高，差异均有统计学意义（P均<0.05）；而A、B、D三组间p-IRS1/IRS1、p-ERK_1/2/ ERK_1/2水平差异均无统计学意义（P>0.05）。结论　VitD-VDR系统减轻高糖诱导的足细胞凋亡，活化胰岛素信号通路，拮抗胰岛素抵抗信号，改善足细胞胰岛素抵抗。

Abstract: 　Objective　To study the protective effects of vitamin D (VitD) on podocyte insulin resistance and its mechanisms. Methods Immortalized mouse podocytes in vitro were randomly divided into 4 groups: podocytes+5mmol/L glucose group (group A); podocytes+5mmol/L glucose+1nmol/L propylene glycol group (group B); podocytes+30mmol/L glucose+1nmol/L propylene glycol group (group C); podocytes+30mmol/L glucose+1nmol/L propylene glycol+1nmol/L VitD group (group D). The percentage of podocyte apoptosis was determined after 48 h of incubation. Podocyte viability was assessed by MTT assay. The mRNA expressions of vitamin D receptor (VDR) and insulin receptor substrate protein 1 (IRS1) in podocyte were detected by RT-PCR. Western blot analysis was performed to measure the protein levels of p-IRS1/IRS, p-Akt/Akt and p-ERK1/2/ERK1/2. Results There were significant differences in apoptosis percentage, viability and the expression of VDR, IRS1, p-ERK1/2 of podoctyes（P<0.05）among 4 groups. There was no difference in p-Akt/Akt expression among 4 groups（P>0.05）. Compared with group A, B , and D, the percentage of podocyte apoptosis in group C was significantly increased, the cell viability was decreased, the expressions of VDR and IRS1 mRNA and p-IRS1 and p-Akt proteins were down-regulated, whereas p-ERK1/2 was up-regulated in group C. The levels of p-IRS1/IRS1, p-Akt/Akt, p-ERK1/2 / ERK1/2 had no statistical differences in group A, B, and D (P>0.05). Conclusions VitD-VDR system alleviates podocyte apoptosis induced by high glucose, and activates insulin signaling pathway and counteracts insulin resistance signal to improve podocyte insulin resistance.

邹敏书, 余健, 聂国明, 罗莉漫, 徐洪涛, 尹晓玲, 周建华. 维生素D对高糖诱导足细胞胰岛素抵抗的保护作用[J]. 临床儿科杂志, 2014, 32(5): 471-475.

ZOU Minshu,YU Jian,NIE Guoming,LUO Liman,XU Hongtao,YIN Xiaoling,ZHOU Jianhua. Protective effect of vitamin D on high glucose-induced podocyte insulin resistance [J]. , 2014, 32(5): 471-475.

参考文献

[1] Welsh GI，Hale LJ，Eremina V，et al. Insulin signaling to the glomerular podocyte is critical for normal kidney function[J]. Cell Metab，2010，12(4)：329-340.
[2] de Zeeuw D，Agarwal R，Amdahl M，et al. Selective vitamin D receptor activation with paricalcitol for reduction of albuminuria in patients with type 2 diabetes (VITAL study)：a randomised controlled trial [J]. Lancet，2010，376(9752)：1543-1551.
[3] 邹敏书，余健，聂国明，等. 帕立骨化醇对糖尿病大鼠肾脏保护作用的机制探讨[J].解放军药学学报，2011，27(6)：476-479.
[4] Ma J，Zhang B，Liu S，et al. 1，25-Dihydroxyvitamin D(3) inhibits podocyte uPAR expression and reduces proteinuria[J]. PLoS One，2013，8(5)：e64912.
[5] Coward RJ，Saleem MA. Podocytes as a target of insulin[J]. Curr Diabetes Rev，2011，7(1)：22-27.　
[6] Copps KD，White MF. Regulation of insulin sensitivity by serine/threonine phosphorylation of insulin receptor substrate proteins IRS1 and IRS2[J]. Diabetologia，2012，55(10)：2565-2582.　
[7] Fornoni A. Proteinuria，the podocyte，and insulin resistance[J]. N Engl J Med，2010，363(21)：2068-2069.　
[8] Yu Q，Gao F，Ma XL. Insulin says NO to cardiovascular disease[J]. Cardiovasc Res，2011，89(3)：516-524.　
[9] Tang YC，Liu CW，Chang HH，et al. Endothelin-1 stimulates resistin gene expression[J]. Endocrinology，2014 ，155（3）：854-864.
[10] Liu HY，Collins QF，Xiong Y，et al. Prolonged treatment of primary hepatocytes with oleate induces insulin resistance through p38 mitogen-activated protein kinase[J]. J Biol Chem，2007，282(19)：14205-14212.　
[11] Du P，Fan B，Han H，et al. NOD2 promotes renal injury by exacerbating inflammation and podocyte insulin resistance in diabetic nephropathy[J]. Kidney Int，2013，84(2)：265-276.　
[12] De Cosmo S，Menzaghi C，Prudente S，et al. Role of insulin resistance in kidney dysfunction： insights into the mechanism and epidemiological evidence[J]. Nephrol Dial Transplant，2013，28(1)：29-36.　
[13] 李海园，刘志伟. 维生素D的免疫调节作用与儿童疾病[J].临床儿科杂志，2012，30（8）：792-794.
[14] Karnchanasorn R，Ou HY，Chiu KC. Plasma 25-hydroxyvitamin D levels are favorably associated with β-cell function[J]. Pancreas，2012，41(6)：863-868.　
[15] von Hurst PR，Stonehouse W，Coad J. Vitamin D supplementation reduces insulin resistance in South Asian women living in New Zealand who are insulin resistant and vitamin D deficient-a randomised，placebo-controlled trial[J]. Br J Nutr，2010，103(4)：549-555.　
[16] Al-Daghri NM，Al-Attas OS，Alokail MS，et al. Hypovitaminosis D associations with adverse metabolic parameters are accentuated in patients with Type 2 diabetes mellitus： a body mass index-independent role of adiponectin? [J].J Endocrinol Invest，2013，36(1)：1-6.　
[17] Marcotorchino J，Gouranton E，Romier B，et al. Vitamin D reduces the inflammatory response and restores glucose uptake in adipocytes[J]. Mol Nutr Food Res，2012，56(12)：1771-1782.　
[18] Wang Y，Deb DK，Zhang Z，et al. Vitamin D receptor signaling in podocytes protects against diabetic nephropathy[J]. J Am Soc Nephrol，2012，23(12)：1977-1986.　
[19] 邹敏书，余健，周建华，等. 活性维生素D及其类似物对足细胞的保护作用[J].临床儿科杂志，2012，30（1）：93-96.
[20] 许桂平，于明香.维生素D与糖尿病[J].复旦学报(医学版).2011，38(1)：80-83.